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Assessment of Mitochondrial Functions and Cell Viability in Renal Cells Overexpressing Protein Kinase C Isozymes
Published on: January 7, 2013
Cellular and molecular studies on cisplatin-induced apoptotic cell death in rat kidney
David Sheikh-Hamad1, William Cacini, Arthur R Buckley
1Departments of Medicine and Renal Pathology, Baylor College of Medicine, Houston, TX 77030, USA.
Abstract:
Using morphological and molecular approaches, we characterized cisplatin-induced cell necrosis and apoptosis in rat kidney. Male Sprague-Dawley rats ( n=5 per group) received a single intraperitoneal injection of either cisplatin (5 mg/kg) or saline, and were killed on day 5. Functionally, cisplatin-treated rats developed polyuric acute renal failure. Morphologically, kidneys of cisplatin-treated rats showed overt tubular necrosis associated with apoptosis in the corticomedullary junction. Cell necrosis was segment-specific and was distributed in radial fashion at the corticomedullary junction. The apoptosis was limited to discrete cells in apparently intact tubules in the vicinity of the necrosed tubules. The apoptotic changes were confirmed by TUNEL (TdT-mediated deoxyuridine triphosphate nick-end labeling) and staining for cleaved caspase-3. Analysis of outer medullary tissue for apoptosis-related molecules by RNase protection assay revealed a significant increase in the expression of pro-apoptotic mRNAs (caspases 1, 2, and 8, and Bax) in cisplatin-treated rats. On the other hand, the expression of mRNA for the anti-apoptotic Bcl-2 did not change, resulting in a decrease in relative ratio of Bcl-2/Bax, and thus favoring apoptosis. The above changes were paralleled by a marked increase in caspase-3 precursor, the executioner protease. Furthermore, these pro-apoptotic molecular changes were associated with a 3-fold increase in the activity of JNK1 in the outer medulla, but not in the cortex, of cisplatin-treated rat kidneys, localizing to the site of maximal apoptosis. Upregulation of JNK1 activity in the outer medulla was not accompanied by changes in the activities of ERK or p38 kinase. In conclusion, these data suggest that cisplatin-induced apoptotic cell death in native kidney may be mediated by cooperative activation of the JNK1 pathway and Bax in the outer medulla.
Insights
Cisplatin causes kidney failure by inducing cell necrosis and apoptosis, particularly in the outer medulla. This process involves the JNK1 pathway and Bax, leading to programmed cell death.
Area of Science:
- Nephrology
- Molecular Biology
- Cell Biology
Background:
- Cisplatin is a widely used chemotherapy agent with known nephrotoxic side effects.
- The precise mechanisms of cisplatin-induced kidney damage, including cell death pathways, require further elucidation.
Purpose of the Study:
- To characterize the morphological and molecular mechanisms of cisplatin-induced cell necrosis and apoptosis in rat kidneys.
- To investigate the role of specific signaling pathways, such as JNK1, in cisplatin nephrotoxicity.
Main Methods:
- Rats received cisplatin or saline, and kidney tissues were analyzed morphologically and molecularly.
- Techniques included TUNEL staining, cleaved caspase-3 staining, RNase protection assay, and kinase activity assays.
Main Results:
- Cisplatin induced acute renal failure, tubular necrosis, and apoptosis in the corticomedullary junction.
- Pro-apoptotic gene expression (caspases 1, 2, 8, Bax) increased, while the Bcl-2/Bax ratio decreased.
- JNK1 activity significantly increased in the outer medulla, correlating with maximal apoptosis.
Conclusions:
- Cisplatin-induced kidney apoptosis is mediated by the JNK1 pathway and Bax in the outer medulla.
- These findings provide insights into the molecular basis of cisplatin nephrotoxicity.
- Targeting the JNK1 pathway could be a potential therapeutic strategy to mitigate cisplatin-induced kidney damage.
